Use for 1,3-propanediol derivatives

ABSTRACT

Use of a compound of formula I  
                 
 
     wherein  
     R 1  is an optionally substituted straight- or branched carbon chain having 12 to 22 carbon atoms which may be optionally interrupted by an optionally substituted phenylene, and  
     each of R 2 , R 3 , R 4  and R 5 , independently, is H or lower alkyl,  
     in free form or in pharmaceutically acceptable salt form,  
     in the prevention or treatment of chronic rejection in a recipient of organ or tissue allo- or xenotransplant, or of acute rejection in a xenograft transplant recipient.

[0001] The present invention relates to a new use for a compound groupcomprising 2-amino-1,3-propanediol derivatives.

[0002] Compounds for use according to the invention are compounds offormula I

[0003] wherein

[0004] R₁ is an optionally substituted straight- or branched carbonchain having 12 to 22 carbon atoms which may be optionally interruptedby an optionally substituted phenylene, and

[0005] each of R₂, R₃, R₄ and R₅, independently, is H or lower alkyl,

[0006] in free form or in pharmaceutically acceptable salt form.

[0007] When the carbon chain as R₁ is substituted, it is preferablysubstituted by halogen, nitro, amino, hydroxy or carboxy. When thecarbon chain is interrupted by an optionally substituted phenylene, thecarbon chain is preferably unsubstituted. When the phenylene moiety issubstituted, it is preferably substituted by halogen, nitro, amino,methoxy, hydroxy or carboxy.

[0008] Such compounds are disclosed in EP-A1-627,406 the relevantdisclosure of which, in particular with respect to the compounds, isincorporated herein by reference.

[0009] Preferred compounds of formula I are those wherein R₁ is astraight or branched, preferably straight, chain alkyl having 13 to 20carbon atoms, optionally substituted by nitro, halogen, amino, hydroxyor carboxy, and, more preferably those wherein R₁ is phenylalkylsubstituted by a straight or branched C₆₋₁₄ alkyl chain optionallysubstituted by halogen and the alkyl moiety is a C₆₋₁₄ alkyl optionallysubstituted by hydroxy. More preferably, R₁ is phenyl-C₁₋₆ alkylsubstituted on the phenyl by a straight or branched, preferablystraight, C₆₋₁₄ alkyl chain. The C₆₋₁₄ alkyl chain may be in ortho, metaor para, preferably in para.

[0010] Preferably each of R₂ to R₅ is H.

[0011] Examples of the pharmaceutically acceptable salts of thecompounds of the formula (I) include salts with inorganic acids, such ashydrochloride, hydrobromide and sulfate, salts with organic acids, suchas acetate, fumarate, maleate, benzoate, citrate, malate,methanesulfonate and benzenesulfonate salts, and when a carboxy group ispresent, salts with metals such as sodium, potassium, calcium andaluminium, salts with amines, such as triethylamine and salts withdibasic amino acids, such as lysine. The compounds and salts of thepresent invention encompass hydrate and solvate forms.

[0012] When the compounds of formula I have one or more asymmetriccenters in the molecule, the present invention is to be understood asembracing the various optical isomers, as well as racemates,diastereoisomers and mixtures thereof are embraced.

[0013] Particularly preferred compounds of formula I are2-amino-2-tetradecyl-1,3-propanediol and especially2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol, (hereinafterCompound A) e.g. in hydrochloride form.

[0014] Compounds of formula I have, on the basis of observed activity,e.g. as described in EP-A1-627,406 been found to be useful e.g. asimmunosuppressant, e.g. in the treatment of acute allograft rejection.

[0015] Organ transplants of liver, kidney, lung and heart are nowregularly performed as treatment for endstage organ disease. Allograftas well as xenograft transplants have been performed. However, becauseof problems with long-term chronic rejection, organ transplantation isnot yet a permanent solution to irreversible organ disease. Chronicrejection, which manifests as progressive and irreversible graftdysfunction, is the leading cause of organ transplant loss, in somecases already after the first postoperative year. The clinical problemof chronic rejection is clear from transplantation survival times; abouthalf of kidney allografts are lost within 5 years after transplantation,and a similar value is observed in patients with a heart allograft.

[0016] Chronic rejection is considered as a multifactorial process inwhich not only the immune reaction towards the graft but also theresponse of the blood vessel wall in the grafted organ to injury(“response-to-injury” reaction) plays a role. The variant of chronicrejection with the worst prognosis is an arteriosclerosis-likealteration, also called transplant vasculopathy graft vessel disease,graft atherosclerosis, transplant coronary disease, etc. This vascularlesion is characterized by migration and proliferation of smooth musclecells under influence of growth factors, that are amongst otherssynthesized by endothelium. It appears to progress also throughrepetitive endothelial injury induced amongst others by host antibody orantigen-antibody complexes, through intimal proliferation andthickening, smooth muscle cell hypertrophy repair, and finally togradual luminal obliteration. Also so-called non-immunological factorslike hypertension, hyperlipidemia, hypercholesterolemia etc. play arole.

[0017] Chronic rejection appears to be inexorable and uncontrollablebecause there is no known effective treatment or prevention modality.Thus, there continues to exist a need for a treatment effective inpreventing, controlling or reversing manifestations of chronic graftvessel diseases.

[0018] In accordance with the present invention, it has now surprisinglybeen found that compounds of formula I in free form or inpharmaceutically acceptable salt form inhibit graft vessel disease andare particularly indicated to prevent or treat chronic rejection in atransplanted organ.

[0019] Furthermore, it has also been found that compounds of formula Iin free form or in pharmaceutically acceptable salt form suppressxenograft rejection. In accordance with the particular findings of thepresent invention, there is provided:

[0020] 1.1. A method of preventing or treating manifestations of chronicrejection, e.g. to avoid, reduce or restrict chronic rejection, in arecipient of organ or tissue allo- or xeno-transplant, e.g. heart, lung,combined heart-lung, liver, kidney or pancreatic transplant, comprisingthe step of administering to said recipient a therapeutically effectiveamount of a compound of formula I in free form or in pharmaceuticallyacceptable salt form;

[0021] 1.2. A method of preventing or treating graft vessel diseases,e.g. transplant vasculopathy, arteriosclerosis or atherosclerosis, in arecipient of organ or tissue allo- or xeno-transplant, e.g. heart, lung,combined heart-lung, liver, kidney or pancreatic transplants, comprisingthe step of administering to said recipient a therapeutically effectiveamount of a compound of formula I in free form or in pharmaceuticallyacceptable salt form;

[0022] In a series of further specific or alternative embodiments, thepresent invention also provides:

[0023] 2. A method of preventing or controlling acute rejection in axenograft transplant recipient, e.g. a patient receiving a heart, lung,combined heart-lung, kidney, liver, bone marrow, pancreatic bowel, skinor corneal xenotransplant, comprising administering to said recipient atherapeutically effective amount of a compound of formula I in free formor in pharmaceutically acceptable salt form.

[0024] As alternative to the above the present invention also provides:

[0025] 3. A compound of formula I in free form or in pharmaceuticallyacceptable salt form for use in any method as defined under 1 or 2above; or

[0026] 4. A compound of formula I in free form or in pharmaceuticallyacceptable salt form for use in the preparation of a pharmaceuticalcomposition for use in any method as defined under 1 or 2 above; or

[0027] 5. A pharmaceutical composition for use in any method as definedunder 1 or 2 above comprising a compound of formula I in free form or inpharmaceutically acceptable salt form together with one or morepharmaceutically acceptable diluents or carriers therefor.

[0028] Utility of the compounds of formula I in free form or inpharmaceutically acceptable salt form in chronic rejection, as well asutility in treating diseases and conditions as hereinabove specified,may be demonstrated in animal tests for example in accordance with themethods hereinafter described, as well as in clinic where e.g. thetransplanted organ or tissue may be submitted to regular biopsy controlsand in the case of heart transplant additionally to ultrasound scanning.

[0029] A. Prevention of Graft Vessel Disease

[0030] Experimental Animals:

[0031] Inbred rat strains DA (RT1^(a), donors) and Lewis (RT1¹,recipients of allografts), weighing between 200 and 350 grams are used.The animals are allowed unrestricted access to food and water before andafter the operation.

[0032] Carotid Artery Transplantation:

[0033] The rats are anaesthetised with isofluorane (Abbott), (4-5% forinduction, 1.5-2% for maintenance) and 300 μg atropin sulphate isinjected subcutaneously following the induction. The left carotid arteryis dissected free. The artery is clamped proximally and distally and asegment of about 7-10 mm is removed. The gap is bridged by an allograftwhich had also been subjected to 45 min cold ischemia. Ethilon 10/osutures are used. Finally the skin is closed with 4/0 sutures. Ifneeded, an Alzet osmotic minipump (Alza Corp. Palo Alto, Calif.) is thenimplanted subcutaneously over the back (alternatively the animals aretreated orally).

[0034] The rats are subject to one of the following treatments: Acompound of formula I alone at the doses of 0.1 to 10 mg/kg, or incombination with Cyclosporin A at the dose of 0.03, 0.3, or 1mg.kg⁻¹.day⁻¹ is administered for 8 weeks either by using Alzet osmoticminipumps implanted subcutaneously or alternatively by oraladministration. At 8 weeks the rats are sacrificed, the carotid arteriesare perfused for 1 min with 0.1 M phosphate buffered saline solution(PBS, pH 7.4) and then for 15 min with 2.5% glutaraldehyde in phosphatebuffer (pH 7.4). The carotid arteries are then excised and stained inGiemsa solution for histological evaluation. Morphometric analysisincludes the measurement of the thickness of the media and intima. Aqualitative analysis of the morphological changes includes a scoring onan 0-3 scale for adventitial infiltration of mononuclear cells andnecrosis (vacuolar degeneration, hypertrophy of cells), the number ofsmooth muscle cells (SMC) nuclei in the media (0-10, <100, >100and >>100 nuclei for scores, 0, 1, 2 and 3 respectively, SMC necrosis(vacuolar degeneration and hypertrophy of SMC) and the intimalinfiltration of mononuclear cells (13).

[0035] In both experiments, the compounds of formula I, particularlyCompound A in hydrochloride form, significantly inhibit graftinfiltration and neointima formation.

[0036] B. In vivo Heart Xenotransplantation (Hamster-to-rat)

[0037] The hamster-into-rat xenograft combination is a so-calleddifficult concordant combination. Rats do not have natural anti-hamsterantibody in sufficient amounts to yield immediate hyperacute rejectionas observed in concordant combinations; however, rejection in untreatedrecipients occurs within 3-4 days, by antibodies in combination withcomplement. This is visualized in histology by destruction of bloodvessels, exsudation and extravasation of erythrocytes, and influx bypolymorphonuclear granulocytes; often there are signs of hemorrhage andthrombosis. Once this rejection has been overcome by effectiveinhibition of antibody synthesis or complement inactivation, a cellularrejection can emerge later on. This is visualized in histology by influxof mononuclear cells, including lymphocytes, lymphoblastoid cells, andmacrophages, and destruction of the myocyte parenchyma. The inhibitionof cellular rejection requires more immuno-suppression than that ofallografts. Congenitally athymic (rnu/rnu) rats lack a competent(thymus-dependent) cellular immune system and generally are unable toreject allografts. Such animals do reject a hamster xenograft within 3-4days in a similar fashion as euthymic rats, indicative that (at leastpart of) anti-hamster antibody synthesis in rats occurs following athymus-independent B-cell response. Such recipients are useful inhamster xenografting to evaluate rejection by thymus-independentantibody-mediated rejection.

[0038] The heart of a Syrian hamster is heterotopically transplanted inthe abdomen of a male Lewis (RT1′) rat with anastomoses between thedonor and recipient's aorta and the donor right pulmonary artery to therecipient's inferior vena cava. The graft is monitored daily bypalpation of the abdomen. Rejection is concluded in case of cessation ofheart beat. Animals are weighed weekly. In the present series ofexperiments, the endpoint is set to 28 days. Animals are subjected toautopsy; apart from the graft, weight and histology is assessed forthymus, spleen, liver, seminal vesicles and testes. Blood is taken andprocessed to serum for the determination of cytolytic anti-hamstererythrocyte antibody and hemolytic complement activity.

[0039] Compounds are dissolved in water and administered daily orally ina volume of 2 ml/kg body weight. Administration of 5-30 mg/kg/day, of acompound of Formula I, e.g. Compound A in hydrochloride form, results inprolonged graft survival, in both athymic and euthymic recipients.

[0040] Daily dosages required in practicing the method of the presentinvention will vary depending upon, for example, the compound of formulaI employed, the host, the mode of administration, the severity of thecondition to be treated, and the optionally concomitantly usedimmunosuppressive drug e.g. CysA. A preferred daily dosage range isabout from 0.03 to 2.5 mg/kg per day, particularly 0.1 to 2.5 mg/kg perday, e.g. 0.5 to 2.5 mg/kg per day as a single dose or in divided doses.Suitable daily dosages for patients are on the order of from e.g. 1 to100 mg p.o. Suitable unit dosage forms for oral administration comprisefrom ca. 1 to 50 mg, usually 5 to 30 mg active ingredient, e.g. CompoundA, e.g. in hydrochloride form, together with one or morepharmaceutically acceptable diluents or carriers therefor. As analternative, the compound of formula I in free form or inpharmaceutically acceptable salt form may also be administered twice orthree times a week, e.g. at a dosage as indicated above.

[0041] The compounds of formula I may be administered by anyconventional route, in particular enterally, e.g. orally, for example inthe form of solutions for drinking, tablets or capsules or parenterally,for example in the form of injectable solutions or suspensions.Pharmaceutical compositions comprising the compounds of formula I may bemanufactured in conventional manner, e.g. as described in EP-A1-627,406.

[0042] The compounds of formula I may be administered as the sole activeingredient or together with other drugs in immunomodulating regimens orother anti-inflammatory agents. For example, the compounds of formula Imay be used in combination with cyclosporins, rapamycins or ascomycins,or their immunosuppressive analogs, e.g. cyclosporin A, cyclosporin G,FK-506, rapamycin, 40-O-(2-hydroxy)ethyl-rapamycin, etc.;corticosteroids; cyclophosphamide; azathioprene; methotrexate;brequinar; leflunomide; mizoribine; mycophenolic acid; mycophenolatemofetil; 15-deoxyspergualine; immuno-suppressive monoclonal antibodies,e.g., monoclonal antibodies to leukocyte receptors, e.g., to MHC, CD2,CD3, CD4, CD7, CD25, CD28, B7, CD40, CD45, or CD58 or to their ligands;or other immunomodulatory compounds, e.g. CTLA4-Ig.

[0043] Where the compounds of formula I are administered in conjunctionwith other immunosuppressive/immunomodulatory therapy, e.g. forpreventing or treating chronic rejection as hereinabove specified,dosages of the co-administered immunosuppressant or immuno-modulatorycompound will of course vary depending on the type of co-drug employed,e.g. whether it is a steroid or a cyclosporin, on the specific drugemployed, on the condition being treated, and so forth. In accordancewith the foregoing the present invention provides in a yet furtheraspect:

[0044] 6. A method as defined above comprising co-administration, e.g.concomitantly or in sequence, of a therapeutically effective amount of acompound of formula I and a second drug substance, said second drugsubstance being an immunosuppressant or immunomodulatory drug, e.g. asset forth above.

[0045] 7. A kit or package for use in any method as defined under 1 or 2above, comprising a compound of formula I, in free form or inpharmaceutically acceptable salt form, with at least one pharmaceuticalcomposition comprising an immunosuppressant or immunomodulatory drug.The kit or package may comprise instructions for its administration.

FORMULATION EXAMPLE Soft Capsules

[0046] Compound of formula I, e.g. Compound A  30 mg Polyethylene glycol300 300 mg Polysorbate 80  20 mg Total 350 mg

[0047] Compounds of formula I in free form or in pharmaceuticallyacceptable salt form are well tolerated at dosages required for use inaccordance with the present invention. For example, the acute LD₅₀is >10 mg/kg p.o. in rats and monkeys.

1. Use of a compound of formula I

wherein R₁ is an optionally substituted straight- or branched carbonchain having 12 to 22 carbon atoms which may be optionally interruptedby an optionally substituted phenylene, and each of R₂, R₃, R₄ and R₅,independently, is H or lower alkyl, in free form or in pharmaceuticallyacceptable salt form, in the preparation of a pharmaceutical compositionfor preventing or treating manifestations of chronic rejection in arecipient of organ or tissue allo- or xenotransplant.
 2. Use of acompound of formula I

wherein R₁ is an optionally substituted straight- or branched carbonchain having 12 to 22 carbon atoms which may be optionally interruptedby an optionally substituted phenylene, and each of R₂, R₃, R₄ and R₅,independently, is H or lower alkyl, in free form or in pharmaceuticallyacceptable salt form, in the preparation of a pharmaceutical compositionfor preventing or treating graft vessel diseases in a recipient of organor tissue allo- or xenotransplant.
 3. Use of a compound of formula I

wherein R₁ is an optionally substituted straight- or branched carbonchain having 12 to 22 carbon atoms which may be optionally interruptedby an optionally substituted phenylene, and each of R₂, R₃, R₄ and R₅,independently, is H or lower alkyl, in free form or in pharmaceuticallyacceptable salt form, in the preparation of a pharmaceutical compositionfor preventing or controlling acute rejection in a xenograft transplantrecipient.
 4. Use of a compound of formula I as defined in claim 1 , infree form or in a pharmaceutically acceptable salt form, for preventingor treating manifestations of chronic rejection in a recipient of organor tissue allo- or xenotransplant.
 5. Use of a compound of formula I asdefined in claim 1 , in free form or in a pharmaceutically acceptablesalt form, for preventing or controlling acute rejection in a xenografttransplant recipient.
 6. Use according to any one of the precedingclaims, wherein the compound of formula I is2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol, in free form or inpharmaceutically acceptable salt form.
 7. A pharmaceutical compositionfor preventing or treating manifestations of chronic rejection in arecipient of organ or tissue allo- or xenotransplant, or for preventingor controlling acute rejection in a xenograft transplant recipient,comprising a compound of formula I as defined in claim 1 , in free formor in a pharmaceutically acceptable salt form, together with one or morepharmaceutically acceptable diluents or carriers therefor.
 8. A kit orpackage for preventing or treating manifestations of chronic rejectionin a recipient of organ or tissue allo- or xenotransplant, or forpreventing or controlling acute rejection in a xenograft transplantrecipient, comprising a compound of formula I, in free form or in apharmaceutically acceptable salt form, with a pharmaceutical compositioncomprising an immunosuppressant or immunomodulatory drug.
 9. Acomposition according to claim 7 or a kit or package according to claim8 , wherein the compound of formula I is2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol, in free form or inpharmaceutically acceptable salt form.
 10. A method of preventing ortreating manifestations of chronic rejection in a recipient of organ ortissue allo- or xenotransplant, or of preventing or controlling acuterejection in a xenograft transplant recipient, comprising the step ofadministering to said recipient a therapeutically effective amount of acompound of formula I as defined in claim 1 in free form or inpharmaceutically acceptable salt form.